JP2018507245A - Brain tumor treatment methods - Google Patents

Abstract

Disclosed herein is a method of treating brain tumors by administering plinabulin. Some embodiments relate to the treatment of polymorphic glioblastomas by administering plinabulin.

Description

Related Art The present application is based on US Provisional Patent Application No. 62 / 129,623 filed on March 6, 2015 and US Provisional Patent Application No. 62 / 249,807 filed on November 2, 2015 (these The disclosure of which is incorporated herein by reference in its entirety.

TECHNICAL FIELD The present invention relates to the fields of chemistry and medicine. More particularly, the present invention relates to a method for treating brain tumors using plinabulin.

  Brain and nervous system cancers are among the most difficult to treat. The prognosis for these cancer patients depends not only on their onset stage but also on the type and location of the tumor. Because there are many types of brain cancer, the average lifespan after onset can be several months or 1-2 years. Treatment mainly consists of surgical excision and radiation therapy. Chemotherapy is also used, but the range of appropriate chemotherapeutic agents is limited, probably because most therapeutic agents do not penetrate the blood-brain barrier sufficient to treat brain tumors. The use of known chemotherapeutic drugs in addition to surgery and radiation rarely extends the survival rate obtained with surgery and radiation alone. Therefore, improved treatment options for brain tumors are needed and there is an urgent unmet medical need in this situation.

  Polymorphic glioblastoma (GBM) is the most common adult primary brain tumor, famous for its mortality and lack of response to current therapies. There has been no substantial improvement in treatment options in recent years, only minimal improvement in the likelihood of survival for GBM patients. For GBM, life expectancy after onset is approximately 6-12 months. In addition, there are no approved drugs for the treatment of metastatic brain tumors with a life expectancy after onset of 4-6 months. Thus, there remains an urgent need for improved treatment for brain cancer.

  Some embodiments relate to a method of treating a brain tumor comprising administering an effective amount of plinabulin to a subject in need thereof.

  Some embodiments relate to a method of inhibiting growth of brain tumor cells comprising contacting the brain tumor cells with plinabulin.

  Some embodiments relate to a method for inducing apoptosis of brain tumor cells comprising contacting the brain tumor cells with plinabulin.

  Some embodiments relate to a method of inhibiting the progression of brain tumor comprising administering an effective amount of plinabulin to a subject in need thereof.

FIGS. 1 a-1 d show a glioblastoma (GBM) proneural genetically modified mouse model (GEMM) that mimics human pathology. FIG. 1a shows a T2 MRI image of human GBM showing peri-tumor edema; FIG. 1b shows a T2 MRI image of mouse GBM showing peri-tumor edema; FIG. 1c shows characteristic pseudo-fenced necrosis and FIG. 1d shows a human micrograph image of GBM H & E staining showing microvascular growth; FIG. 1d shows a mouse micrograph image of GBM H & E staining showing characteristic pseudofence-like necrosis and microvascular growth. 2A and 2B show T2-weighted MRI images. FIG. 2A shows the tumor size of mice with PDGF-induced glioma treated with vehicle, temozolomide or fractionated radiation. 2A and 2B show T2-weighted MRI images. FIG. 2B shows the survival rate of mice with PDGF-induced glioma treated with vehicle, temozolomide or fractionated radiation. Figure 3 shows the survival of mice with glioblastoma tumors treated with control and plinabulin. Figure 6 shows the survival of mice with PDGF-induced gliomas characterized by the combination of plinabulin, temozolomide, and radiation and the expression of KRAS mutations treated with temozolomide and radiation combination.

  Plinabulin, (3Z, 6Z) -3-benzylidene-6-{[5- (2-methyl-2-propanyl) -1H-imidazol-4-yl] methylene} -2,5-piperazinedione is a natural compound phenyl A synthetic analog of ahistine. Plinabulin can be readily prepared according to the methods and procedures described in US Pat. Nos. 7,064,201 and 7,919,497, which are hereby incorporated by reference in their entirety. . Some embodiments are for brain tumor treatment, including but not limited to metastatic brain tumors, anaplastic astrocytoma, polymorphic glioblastoma, oligodendroglioma, ependymoma, and mixed glioma Related to the use of plinabulin. Some embodiments relate to the use of plinabulin for inhibiting growth of brain tumor cells using plinabulin. Some embodiments relate to the use of plinabulin for inducing apoptosis of brain tumor cells using plinabulin. Some embodiments relate to the use of plinabulin for inhibition of brain tumor progression. Some embodiments relate to the use of plinabulin in combination with an additional therapeutic agent or radiation for inhibition of brain tumor progression.

Definitions Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. All patents, applications, published applications, and other publications are incorporated by reference in their entirety. Where there are multiple definitions of terms herein, those in this section apply unless otherwise specified.

  As used herein, a “subject” is a human or non-human mammal, such as a dog, cat, mouse, rat, cow, sheep, pig, goat, non-human primate or bird, such as a chicken, and Means other vertebrates or invertebrates.

  The term “mammal” is used in its normal biological sense. Accordingly, details include primates including monkeys (chimpanzees, apes, monkeys) and humans, cows, horses, sheep, goats, pigs, rabbits, dogs, cats, rodents, rats, mice, guinea pigs, and the like. However, it is not limited to this.

  As used herein, an “effective amount” or “therapeutically effective amount” has the effect of alleviating one or more symptoms of a disease or condition to some extent, or reducing the likelihood of its onset, and a disease or condition. Represents the amount of therapeutic agent, including healing.

As used herein, “treat”, “treatment”, or “treating” refers to administering a compound or pharmaceutical composition to a subject for prophylactic and / or therapeutic purposes.
The term “prophylactic treatment” refers to treating a subject who has not yet shown signs of a disease or condition but is susceptible to or otherwise at risk of a particular disease or condition, whereby the treatment Represents a reduction in the likelihood of developing the disease or condition. The term “therapeutic treatment” refers to administration therapy to a subject already suffering from a disease or condition.

The term “pharmaceutically acceptable salt” refers to a salt that retains the biological effectiveness and properties of the compound and which is not biologically or otherwise harmful to pharmaceutical use. In most cases, the compounds disclosed herein can form acid and / or base salts by the presence of amino and / or carboxyl groups or groups similar thereto. Pharmaceutically acceptable acid addition salts can be formed with inorganic and organic acids. Examples of inorganic acids from which salts can be derived include hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid and the like. Examples of organic acids from which salts can be derived include acetic acid, propionic acid, glycolic acid, pyruvic acid, oxalic acid, maleic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, citric acid, benzoic acid, cinnamic acid, mandel Examples include acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like. Pharmaceutically acceptable salts can also be produced using inorganic and organic bases. Inorganic bases from which salts can be derived include, for example, bases containing sodium, potassium, lithium, ammonium, calcium, magnesium, iron, zinc, copper, manganese, aluminum, etc .; particularly preferred are ammonium salts, potassium Salt, sodium salt, calcium salt and magnesium salt. In some embodiments, treatment of a compound disclosed herein with an inorganic base results in the compound losing its reactive hydrogen and an inorganic cation such as Li ⁺ , Na ⁺ , K ⁺ , Mg ²⁺ and Ca ^2+. To obtain a salt form comprising Organic bases from which salts can be derived include, for example, primary, secondary, and tertiary amines, substituted amines including natural substituted amines, cyclic amines, basic ion exchange resins, etc., in particular, isopropylamine, trimethylamine, Examples include diethylamine, triethylamine, tripropylamine, and ethanolamine. As described in Johnston et al., WO 87/05297, published Sep. 11, 1987, which is incorporated herein by reference in its entirety, many such salts are It is known in the art.

  In some embodiments, the composition can further comprise one or more pharmaceutically acceptable diluents. In some embodiments, the pharmaceutically acceptable diluent can comprise Kollifor® (polyethylene glycol (15) -hydroxystearate). In some embodiments, the pharmaceutically acceptable diluent can include propylene glycol. In some embodiments, pharmaceutically acceptable diluents can include corifol and propylene glycol. In some embodiments, the pharmaceutically acceptable diluent can include collifol and propylene glycol, the collifol is about 40% by weight relative to the total weight of the diluent, and the propylene glycol is about 60% by weight. In some embodiments, the composition can further comprise one or more other pharmaceutically acceptable excipients.

  Standard pharmaceutical formulation techniques are described herein, including those described in Remington's The Science and Practice of Pharmacy, 21st Ed., Lippincott Williams & Wilkins (2005), which is incorporated herein by reference in its entirety. Can be used to produce the pharmaceutical composition described in 1. above. Thus, some embodiments are: (a) a safe and therapeutically effective amount of plinabulin or a pharmaceutically acceptable salt thereof; and (b) a pharmaceutically acceptable carrier, diluent, excipient or A pharmaceutical composition comprising the combination.

Other embodiments include co-administration of plinabulin and an additional therapeutic agent in separate or the same composition. Accordingly, some embodiments include (a) a safe and therapeutically effective amount of plinabulin or a pharmaceutically acceptable salt thereof and (b) a pharmaceutically acceptable carrier, diluent, excipient or its A first pharmaceutical composition comprising a combination; and (a) a safe and therapeutically effective amount of an additional therapeutic agent and (b) a second comprising a pharmaceutically acceptable carrier, diluent, excipient or combination thereof Including a pharmaceutical composition. Some embodiments include (a) a safe and therapeutically effective amount of plinabulin or a pharmaceutically acceptable salt thereof; (b) a safe and therapeutically effective amount of an additional therapeutic agent; and (c) an agent. Pharmaceutical compositions comprising a pharmaceutically acceptable carrier, diluent, excipient or combination thereof.

  Administration of the pharmaceutical compositions described herein is not limited to oral, sublingual, buccal, subcutaneous, intravenous, intranasal, topical, transdermal, intradermal, intraperitoneal, intramuscular, intrapulmonary Any of the accepted methods of administration of agents that serve similar utilities, including intravaginal, rectal, or intraocular. Oral and parenteral administration are routine in treating the symptoms that are the subject of the preferred embodiment.

The term “pharmaceutically acceptable carrier” or “pharmaceutically acceptable excipient” refers to any and all solvents, dispersion media, coatings, antibacterial and antifungal agents, isotonic and absorption delaying Includes agents and the like. The use of such media and agents for pharmaceutical agents is well known in the art. Except where conventional media or drugs are incompatible with the active ingredient, use in therapeutic compositions is expected. In addition, various adjuvants as commonly used in the art may be included. For discussion of the inclusion of various ingredients in pharmaceutical compositions, see, for example, Gilman et al. (Eds.) (1990); Goodman and Gilman's: The Pharmacological Basis of
Therapeutics, 8th Ed., Pergamon Press (incorporated herein by reference in its entirety).

  Some examples of substances that can serve as pharmaceutically acceptable carriers or components thereof include sugars such as lactose, glucose and sucrose; starches such as corn starch and potato starch; sodium carboxymethylcellulose, ethylcellulose, and methylcellulose Such as cellulose and its derivatives; tragacanth powder; malt; gelatin; talc; solid lubricants such as stearic acid and magnesium stearate; calcium sulfate; vegetable oils such as peanut oil, cottonseed oil, sesame oil, olive oil, corn oil and cacao oil; Polyols such as glycol, glycerin, sorbitol, mannitol and polyethylene glycol; alginic acid; emulsifier such as tween; wetting agent such as sodium lauryl sulfate Coloring agents; a and phosphate buffer; flavoring; tableting agents, stabilizers; antioxidants; preservatives; pyrogen-free water; isotonic saline.

  The compositions described herein are preferably provided in unit dosage forms. As used herein, a “unit dosage form” is a composition containing an amount of a compound or composition suitable for administration to an animal, preferably a mammalian subject, in a single administration, according to appropriate medical practice. It is. However, single or unit dosage forms do not imply that the dosage form is administered once a day or once per course of treatment. Such dosage forms may be administered once, twice, three times or more daily and may be administered by infusion over a period of time (eg, from about 30 minutes to about 2-6 hours). Or as a continuous infusion, and a single dose is not specifically excluded, but may be given more than once during the course of treatment. One skilled in the art will recognize that the dosage form is not particularly predictive of the entire course of treatment, and that such decisions are left to the skilled artisan rather than the dosage form.

The above useful compositions are available for various routes of administration, eg, oral, sublingual, buccal, nasal, rectal, topical (including transdermal and intradermal), ocular, intracerebral, intracranial, intrathecal, arterial. It can be in any form suitable for internal, intravenous, intramuscular, or other parenteral routes of administration. One skilled in the art will recognize that oral and nasal compositions include compositions that are administered by inhalation and are made by available methods. Depending on the specific route of administration desired, various pharmaceutically acceptable carriers well known in the art may be used. Pharmaceutically acceptable carriers include, for example, solid or liquid fillers, diluents, hydrotropic agents, surfactants, and encapsulating materials.
If desired, pharmaceutically active substances that do not substantially interfere with the activity of the compound or composition may be included. The amount of carrier used with the compound or composition is sufficient to provide a practical amount of the substance to be administered per unit dose of the compound. Techniques and compositions for formulating dosage forms useful in the methods described herein are described in the following references (all incorporated herein by reference): Modern Pharmaceutics, 4th Ed. , Chapters 9 and 10 (Banker & Rhodes, editors, 2002); Lieberman et al., Pharmaceutical Dosage Forms: Tablets (1989); and Ansel, Introduction to Pharmaceutical Dosage Forms 8th Edition (2004).

  A variety of oral dosage forms can be used including solid forms such as tablets, capsules (eg, liquid gel capsules and solid gel capsules), granules and mixed powders. Contains suitable binders, lubricants, diluents, disintegrants, colorants, flavoring agents, glidants, and melting agents, compressed tablets, powder tablets, enteric tablets, dragee tablets, film-coated tablets, or multiple compression It can be a tablet. Liquid oral dosage forms include appropriate solvents, preservatives, emulsifiers, suspending agents, diluents, sweeteners, melting agents, colorants and flavoring agents, aqueous solutions, emulsions, suspensions, non-foaming Liquids and / or suspensions reconstituted from expandable granules, and effervescent preparations reconstituted from effervescent granules.

  Suitable pharmaceutically acceptable carriers for dosage unit formulations for oral administration are well known in the art. Tablets are usually inert diluents such as calcium carbonate, sodium carbonate, mannitol, lactose and cellulose; binders such as starch, gelatin and sucrose; disintegrants such as starch, alginic acid and croscarmellose; magnesium stearate, Contains conventional pharmaceutically compatible adjuvants as lubricants such as stearic acid and talc. Glidants such as silicon dioxide can be used to improve the flow characteristics of the powder mixture. Colorants such as FD & C dyes can be used for appearance. Sweetening and flavoring agents such as aspartame, saccharin, menthol, peppermint, and fruit flavors are useful adjuvants for chewable tablets. Capsules typically include one or more solid diluents disclosed above. The choice of carrier component depends on minor secondary considerations, such as taste, cost, and shelf life, and can be readily manufactured by one skilled in the art.

  Examples of the oral composition include solutions, emulsions, suspensions and the like. Suitable pharmaceutically acceptable carriers for the formulation of such compositions are well known in the art. Typical ingredients for carriers for syrups, elixirs, emulsions and suspensions include ethanol, glycerol, propylene glycol, liquid sucrose, sorbitol and water. For suspending agents, typical suspending agents include methylcellulose, sodium carboxymethylcellulose, AVICEL RC-591, tragacanth and sodium alginate; typical wetting agents include lectin and polysorbate 80; Preservatives include methyl paraben and sodium benzoate. Oral liquid compositions may contain one or more ingredients such as the sweeteners, flavors and colorants disclosed above.

  Such compositions are typically used with pH or time-dependent coatings so that the subject composition is released in the gastrointestinal tract in the vicinity of the desired topical application or at various times when the desired effect is spread. You may coat by the method. Such dosage forms typically include, but are not limited to, one or more of cellulose acetate phthalate, polyvinyl acetate phthalate, hydroxypropylmethylcellulose phthalate, ethylcellulose, Eudragit coating, wax and shellac. Not.

  The composition described herein may contain other medicinal ingredients as necessary.

Other compositions useful for achieving systemic delivery of the subject compounds include sublingual, buccal and nasal dosage forms. Such compositions typically comprise a soluble filler such as sucrose, sorbitol and mannitol; and one or more binders such as acacia, crystalline cellulose, carboxymethylcellulose and hydroxypropylmethylcellulose. Glidants, lubricants, sweeteners, colorants, antioxidants and perfumes as disclosed above may also be included.

  A liquid composition formulated for topical ophthalmic use is formulated for topical administration to the eye. Sometimes formulation considerations (eg drug stability) may not require much optimal comfort, but comfort may be maximized as much as possible. If comfort cannot be maximized, the liquid may be formulated to be acceptable to a patient for topical ophthalmic use. In addition, the ophthalmically acceptable liquid may be packaged for a single use or may contain preservatives to prevent contamination over multiple uses.

  For ophthalmic use, solutions or drugs are often formulated using saline as the primary vehicle. Preferably, the ophthalmic solution may be maintained at a comfortable pH using a suitable buffer system. The formulations may also contain conventional pharmaceutically acceptable preservatives, stabilizers and surfactants.

  Preservatives that may be used in the pharmaceutical compositions disclosed herein include, but are not limited to, benzalkonium chloride, PHMB, chlorobutanol, thimerosal, phenylmercuric acetate, and phenylmercuric nitrate. . A useful surfactant is, for example, Tween 80. Similarly, a variety of useful media may be used in the ophthalmic formulations disclosed herein. These media include, but are not limited to, polyvinyl alcohol, povidone, hydroxypropylmethylcellulose, poloxamer, carboxymethylcellulose, hydroxyethylcellulose and purified water.

  Isotonic modifiers may be added as needed or convenient. Isotonic modifiers include, but are not limited to, salts, particularly sodium chloride, potassium chloride, mannitol and glycerin, or other suitable ophthalmically acceptable isotonic modifiers.

  Various buffers and means for adjusting pH may be used as long as the resulting formulation is ophthalmically acceptable. For many compositions, the pH will be 4-9. Accordingly, examples of the buffer include an acetate buffer, a citrate buffer, a phosphate buffer, and a borate buffer. Acids or bases may be used to adjust the pH of these formulations as needed.

  Ophthalmically acceptable antioxidants include, but are not limited to, sodium metabisulfite, sodium thiosulfate, acetylcysteine, butylated hydroxyanisole and butylated hydroxytoluene.

  Another excipient component that may be included in the ophthalmic formulation is a chelating agent. A useful chelating agent is disodium edetate (EDTA), although other chelating agents may be used instead or in combination.

  For topical use, creams, ointments, gels, solutions or suspensions, etc., containing the compositions disclosed herein are used. Topical formulations may generally consist of a pharmaceutical carrier, a co-solvent, an emulsifier, a penetration enhancer, a preservative system, and an emollient.

For intravenous administration, the compositions described herein may be dissolved or dispersed in a pharmaceutically acceptable diluent such as saline or dextrose. Appropriate excipients may be included to achieve the desired pH, including but not limited to NaOH, sodium carbonate, sodium acetate, HCl, and citric acid. In various embodiments, the pH of the final composition is in the range of 2-8, or preferably 4-7. Antioxidant excipients may include sodium bisulfite, acetone-sodium bisulfite, sodium formaldehyde sulfoxylate, thiourea, and EDTA. Other non-limiting examples of suitable excipients found in the final intravenous composition include sodium or potassium phosphate, citric acid, tartaric acid, gelatin, and carbohydrates such as glucose, mannitol, and dextran. Can be. Further acceptable excipients are Powell, et al., Compendium of Excipients for Parenteral Formulations, PDA J Pharm Sci and Tech 1998, 52 238-311 and Nema et al., Excipients and Their Role in Approved Injectable Products: Current Usage and Future Directions, PDA J Pharm Sci and Tech 2011, 65 287-332, both of which are incorporated herein by reference in their entirety. Containing antibacterial agents may achieve bacteriostatic or fungistatic solutions, including phenylmercury nitrate, thimerosal nitrate, benzethonium chloride, benzalkonium chloride, phenol, cresol, and chlorobutanol. It is not limited.

  Intravenous compositions may be provided to the healthcare provider in another solid form that is reconstituted with a suitable diluent, such as sterile water, saline, or aqueous dextrose, immediately prior to administration. In other embodiments, the composition is provided in a solution ready for parenteral administration. In yet other embodiments, the composition is provided in a solution that is further diluted prior to administration. In embodiments that include administering a combination of the compositions described herein and another agent, the combination may be provided to the healthcare provider as a mixture, or the healthcare provider may administer the two agents prior to administration. Or the two drugs may be administered separately.

The actual dosage of the active compounds described herein will depend on the particular compound and the condition being treated; the selection of the appropriate dosage is well within the knowledge of the skilled artisan. In some embodiments, Purinaburin or a single dose of the other therapeutic agents, from about 5 mg / ^{m 2} ~ about 150 mg / ^{m 2} of body surface area, about 5 mg / ^{m 2} ~ about 100 mg / ^{m 2} of body surface area, body About 10 mg / m ² to about 100 mg / m ² of body surface area, about 10 mg / m ² to about 80 mg / m ² of body surface area, about 10 mg / m ² to about 50 mg / m ² of body surface area, about 10 mg / m ² of body surface area m ² to about 40 mg / m ² , body surface area of about 10 mg / m ² to about 30 mg / m ² , body surface area of about 13.5 mg / m ² to about 100 mg / m ² , body surface area of about 13.5 mg / m ^{2 2} to about 80 mg / m ² , body surface area of about 13.5 mg / m ² to about 50 mg / m ² , body surface area of about 13.5 mg / m ² to about 40 mg / m ² , body surface area of about 13.5 mg / m ² m ² to about 30 mg / m ² , body surface area of about 15 mg / m ² to about 80 mg / m ² , body surface area of about 15 mg / m ² to about 50 mg / m ² , body surface area of about 15 mg / m ² to about 30 mg / m ² . In some embodiments, a single dose of plinabulin or other therapeutic agent can be about 13.5 mg / m ² to about 30 mg / m ² of body surface area. In some embodiments, a single dose of plinabulin or other therapeutic agent is about 5 mg / m ² , about 10 mg / m ² , about 12.5 mg / m ² , about 13.5 mg / m ^{2 of} body surface area, about 15 mg / ^{m 2,} about 17.5 mg / ^{m 2,} about 20 mg / ^{m 2,} about 22.5 mg / ^{m 2,} about 25 mg / ^{m 2,} about 27.5 mg / ^{m 2,} about 30 mg / ^{m 2,} about 40mg / M ² , about 50 mg / m ² , about 60 mg / m ² , about 70 mg / m ² , about 80 mg / m ² , about 90 mg / m ² , or about 100 mg / m ² .

In some embodiments, a single dose of plinabulin or other therapeutic agent is about 5 mg to about 300 mg, about 5 mg to about 200 mg, about 7.5 mg to about 200 mg, about 10 mg to about 100 mg, about 15 mg to about 100 mg. About 20 mg to about 100 mg, about 30 mg to about 100 mg, about 40 mg to about 100 mg, about 10 mg to about 80 mg, about 15 mg to about 80 mg, about 20 mg to about 80 mg, about 30 mg to about 80 mg, about 40 mg to about 80 mg, about It can be 10 mg to about 60 mg, about 15 mg to about 60 mg, about 20 mg to about 60 mg, about 30 mg to about 60 mg, or about 40 mg to about 60 mg. In some embodiments, a single dose of plinabulin or other therapeutic agent can be about 20 mg to about 60 mg, about 27 mg to about 60 mg, about 20 mg to about 45 mg, about 27 mg to about 45 mg. In some embodiments, a single dose of plinabulin or other therapeutic agent is about 5 mg, about 10 mg, about 12.5 mg, about 13.5 mg, about 15 mg, about 17.5 mg, about 20 mg, about 22.5 mg. , About 25 mg, about 27 mg, about 30 mg, about 40 mg, about 50 mg, about 60 mg, about 70 mg, about 80 mg, about 90 mg, about 100 mg, about 125 mg, about 150 mg, or about 200 mg.

  As long as the tumor remains in control and the regimen is clinically tolerated, the duration of administration can be a multiple week treatment cycle. In some embodiments, a single dose of plinabulin or other therapeutic agent can be administered once a week, preferably once on each of days 1 and 8 of a 3 week (21 day) treatment cycle. In some embodiments, a single dose of plinabulin or other therapeutic agent is administered once a week, twice a week, three times a week, four times a week, five times a week, six times a week, or one week for two weeks. It can be administered daily during a 3-week, 4-week, or 5-week treatment cycle. It can be administered on the same day or different days of each week of the treatment cycle.

  As long as the regimen is clinically tolerated, the treatment cycle can be repeated. In some embodiments, the treatment cycle is repeated n times, where n is an integer in the range of 2-30. In some embodiments, n is 2, 3, 4, 5, 6, 7, 8, 9, or 10. In some embodiments, a new treatment cycle can occur immediately after the completion of the previous treatment cycle. In some embodiments, a new treatment cycle can occur at some time after completion of the previous treatment cycle.

  In some embodiments, the compositions described herein can be used in combination with other therapeutic agents. In some embodiments, the compositions described herein can be administered or used in combination with treatments such as chemotherapy, radiation therapy, and biological therapy.

Methods of Treatment Some embodiments relate to methods of treating brain tumors, the methods comprising administering an effective amount of plinabulin to a subject in need thereof.

  In some embodiments, the brain tumor is selected from metastatic brain tumors, anaplastic astrocytoma, polymorphic glioblastoma, oligodendroglioma, ependymoma, meningioma, mixed glioma, and combinations thereof be able to. In some embodiments, the brain tumor is a polymorphic glioblastoma. In some embodiments, the brain tumor is a metastatic brain tumor.

  In some embodiments, the brain tumor is anaplastic astrocytoma, central neuroma, choroid plexus cancer, choroid plexus papilloma, choroid plexus tumor, germ dysplastic neuroepithelial tumor, ependymoma, fibrotic Astrocytoma, giant cell glioblastoma, glioblastoma multiforme, cerebral glioma, gliosarcoma, perivascular cell tumor, medulloblastoma, medullary epithelioma, meningeal carcinomatosis, neuroblastoma, Neurocytoma, oligodendroglioma, oligodendroglioma, optic nerve sheath meningioma, childhood ependymoma, ciliary cell astrocytoma, pineoblastoma, pineal cell tumor, pleomorphic Differentiated neuroblastoma, pleomorphic yellow astrocytoma, primary central nervous system lymphoma, sphenoid wing meningioma, epiphyseal giant cell astrocytoma, epididymal, central nervous system myeloma, And trilateral retinoblastoma.

In some embodiments, the method can include administering an additional therapeutic agent. In some embodiments, the additional therapeutic agent is temozolomide, bevicizumab, everolimus, carmustine, lomustine, procarbazine, vincristine, irinotecan, cisplatin, carboplatin, methotrexate, etoposide v, blastomycin It can be actinomycin, cyclophosphamide, or ifosfamide. In some embodiments, the additional therapeutic agent can be temozolomide. In some embodiments, the additional therapeutic agent can be lomustine.

  In some embodiments, the method can further include subjecting the subject to radiation therapy. In some embodiments, radiation therapy can be whole brain radiation, fractionated radiation therapy, and radiosurgery.

  In some embodiments, the brain tumor is characterized by the expression of a mutant form of KRAS. In some embodiments, the brain tumor is characterized by the expression of a mutated gene that is not KRAS.

  In some embodiments, the methods described herein can further comprise identifying a patient having a cancer characterized by expressing a mutant form of KRAS. In some embodiments, the methods described herein can further comprise identifying a patient having a cancer characterized by expressing a wild type of KRAS. In some embodiments, identification of a patient can include determining whether the patient has a KRAS mutation. Some embodiments relate to a method of treating cancer in a patient identified as having a KRAS mutation, the method comprising administering to the patient a pharmaceutically effective amount of plinabulin, the patient comprising: (Ii) collecting a sample from the patient; (ii) isolating DNA from the sample; (iii) amplifying the KRAS gene or fragment thereof of the isolated DNA; and (iv) whether there is a mutation in the amplified KRAS gene By detecting whether the patient has a cancer characterized by a KRAS mutation is identified.

  Some embodiments relate to a method of inhibiting the growth of brain tumor cells, the method comprising contacting the brain tumor cells with plinabulin. In some embodiments, the contacting comprises administering an effective amount of plinabulin to a subject having brain tumor cells. In some embodiments, the brain tumor is a polymorphic glioblastoma.

  Some embodiments relate to a method of inducing apoptosis of brain tumor cells, the method comprising contacting the brain tumor cells with plinabulin. In some embodiments, the contacting comprises administering an effective amount of plinabulin to a subject having brain tumor cells. In some embodiments, the brain tumor is a polymorphic glioblastoma.

  Some embodiments relate to a method of inhibiting the progression of brain tumors, the method comprising administering an effective amount of plinabulin to a subject in need thereof.

  In order to further illustrate the present invention, the following examples are set forth. Of course, the examples should not be construed as specifically limiting the invention. Variations of these embodiments within the scope of the claims are within the scope of those skilled in the art, and are considered to be within the scope of the description and claimed invention of the specification. Readers skilled in the art using this disclosure and those skilled in the art can prepare and use the present invention without comprehensive examples.

Example 1
The mouse model of glioma used was a PDGF-induced GEMM of glioma that mimics the proneural molecular subgroup of glioblastoma (GBM). This model is based on somatic cell-specific gene transfer; the replicable ALV splice acceptor (RCAS) retroviral system allows the infusion of specific gene mutations within a tightly regulated window of cell type-specific differentiation. It was possible. The RCAS / tv-a system used RCAS retroviral vectors to infect mice that had been genetically modified to express the RCAS receptor (tv-a) in specific cell populations. Here, glioma was generated by RCAS-mediated introduction of PDGF into nestin-expressing cells in the brain. Nestin was expressed in stem / progenitor cell populations in the brain and was shown to be a marker of cancer stem cells in the perivascular (PVN) region in both human and mouse brain tumors. PDGF-induced gliomas developed with full penetrance when combined with Ink4a-arf − / − deletion by 4-5 weeks after infection. These tumors closely mimic the GBM “proneural” subtype in which CDKN2A (encoding both p16INK4A and p14ARF) deletion was observed in 56% of “proneural” human gliomas. Tumor cell structures that define human gliomas such as Scherrel structure, microvessel growth and pseudofenced necrosis were reconstituted in this GEMM as shown in FIGS. Specifically, FIG. 1a shows that the T2 MRI image of human GBM shows peritumor edema; FIG. 1b shows that the T2 MRI image of mouse GBM shows peritumor edema; FIG. FIG. 1d shows a human micrograph image of H & E staining of GBM with characteristic pseudo-barrel necrosis and microvascular growth; FIG. 1d shows a mouse micrograph of H & E staining of GBM with characteristic pseudo-barric necrosis and microvascular growth Images are shown.

  Glioma cells migrated along the white matter track, surrounded neurons and blood vessels, and accumulated at the edge of the brain in the subpuffy space. In this regard, the glioma PDGF-derived GEMM closely resembles PVN-GBM and represents an excellent experimental system that defines the interaction between tumor cells and non-neoplastic cells in the intratumoral microenvironment.

  A glioma PDGF induction model was used to determine the response to radiation and temozolomide as shown in FIGS. 2A and 2B. FIG. 2A shows the tumor size of mice with PDGF-induced glioma treated with vehicle, temozolomide or split radiation; FIG. 2B shows the survival rate of mice with PDGF-induced glioma treated with vehicle, temozolomide or split radiation. Show.

  Glioma-bearing mice were identified by symptoms and confirmed by T2-weighted MRI. These mice were treated with either vehicle, 25 mg / kg temozolomide daily for 12 days, or split radiation (total 20 Gy) at a dose of 2 Gy (grey) daily for 5 days every 2 weeks. The two top images in FIG. 2A show untreated (vehicle) tumor growth; in contrast, temozolomide treated and irradiated tumors were volume reduced over that same period. These tumors recurred after treatment, and all animals died of recurrent tumors as shown by the survival curves of these corresponding mouse cohorts. Data were: 1) tested in this mouse model, 2) their therapeutic effect on mouse survival was very similar to the human condition, 3) all mice died of disease, and 4) of these mouse cohorts The relatively same results demonstrated that it supported the use of this experimental paradigm to detect survival differences in this study.

Mice with PDGF-induced glioma using RCAS / tv-a were generated. Mice are transgenic with RCAS receptor (tv-a) expression from the nestin promoter and background of ink4a / arf − / − and lox-stop-lox luciferase, with RCAS-PDGF, or RCAS-PDGF and Infections were combined with RCAS-KRAS expressing the G12D mutant KRAS. The resulting tumors developed within the first 4-5 weeks in this background for PDGF alone and about one week earlier for tumors resulting from the combination of PDGF and RCAS. Tumors have GBM histological features and can be identified using symptoms of lethargy and poor grooming, MRI scans using T2-weighted sequences, or bioluminescence imaging using an IVIS system. For radiation therapy, mice were treated with 10 Gy daily for a single dose on the temporal side. This treatment prolonged the survival median of the cohort of GBM-bearing mice by about 3 weeks, as shown in FIG. 2B. These treated mice began to gain weight and showed improvement in symptoms within a few days, and their MRI image features showed stabilization or reduction in tumor size, after which they relapsed and died.

Plinabulin was tested on mice with PDGF-induced glioma expressing the G12D mutant KRAS. 4-6 week old nestin-tv-a / ink4a-arf-/-mice were anesthetized with isoflurane and injected with RCAS-PDGF-B-HA, RCAS-KRAS transfected into Df-1 cells. Mice were injected with 1 microliter of a 1: 1 mixture of 2 × 10 ⁵ RCAS-PDGF-B-HA / RCAS-KRAS using a stereotaxic frame with a 26 gauge needle attached to a Hamilton syringe. Cells were injected into the right frontal lobe but adjusted to a cruciform suture of 1.75 mm, lateral -0.5 mm, and depth of 2 mm. Mice were carefully monitored for weight loss and were subjected to testing if they decreased> 0.3 grams over a total of 2 consecutive days or showed external signs of tumor. In the KRAS group, mice were injected intraperitoneally with plinabulin 7.5 mg / kg twice a week for 10 weeks. In the control group, mice were injected with plinabulin diluent alone (40 wt% corifol and 60 wt% propylene glycol). Mice were monitored for lethargy, stooped posture, loss of appetite, external signs of tumor growth, excitement, weight loss and overall growth impairment. Mice were killed if they lost more than 20% of their body weight, mobility, powerlessness, or a weight of 14 grams / male and less than 12 grams for a male. Mice were sacrificed with CO ₂ ; brains were collected and stored overnight in 10% neutral buffered formalin, then replaced with Flex 80 and stored at 4 ° C.

  FIG. 3 shows the survival rate of mice with glioblastoma containing the G12D Kras mutation. As shown in FIG. 3, mice with the glioblastoma PDGF induction model generally had significantly better survival in the plinabulin-treated group when compared to the control group (p = 0.001).

Example 2
Mice with PDGF-induced glioma expressing the G12D mutant KRAS were prepared using the procedure described in Example 1 and used in this experiment. 4-6 week old nestin-tv-a / ink4a-arf-/-mice were anesthetized with isoflurane and injected with RCAS-PDGF-B-HA, RCAS-KRAS transfected into Df-1 cells. Mice were injected with 1 microliter of a 1: 1 mixture of 2 × 10 ⁵ RCAS-PDGF-B-HA / RCAS-KRAS using a stereotaxic frame with a 26 gauge needle attached to a Hamilton syringe. Cells were injected into the right frontal lobe but adjusted to a cruciform suture of 1.75 mm, lateral -0.5 mm, and depth of 2 mm. Mice were carefully monitored for weight loss and were subjected to testing if they decreased> 0.3 grams over a total of 2 consecutive days or showed external signs of tumor.

Mice were placed in two test groups. One group was treated with a combination of temozolomide (TMZ), radiation and plinabulin: irradiated with 10 gy × 1 and intraperitoneally twice weekly for 10 weeks Monday and Thursday with plinabulin 7.5 mg / kg in TMZ and plinabulin dilutions. Administered. The other group, the control group, was treated with a combination of TMZ and radiation: irradiated with 10 gy × 1, and TMZ was administered intraperitoneally twice a week for 10 weeks Monday and Thursday. Mice were monitored for lethargy, stooped posture, loss of appetite, external signs of tumor growth, excitement, weight loss and overall growth impairment. Mice were killed if they lost more than 20% of their body weight, mobility, powerlessness, or a weight of 14 grams / male and less than 12 grams for a male. Mice were sacrificed with CO ₂ ; brains were collected and O / N stored in 10% neutral buffered formalin, then replaced with Flex 80 and stored at 4 ° C. As shown in FIG. 4, mice with the PDGF induction model of glioblastoma generally had significantly better survival in the TMZ + radiotherapy group when compared to the control group receiving plinabulin + TMZ + radiation (p = 0.0149).

Claims (13)

  A method of treating a brain tumor comprising administering an effective amount of plinabulin to a subject in need thereof.   The method of claim 1, wherein the brain tumor is a metastatic brain tumor, an anaplastic astrocytoma, a pleomorphic glioblastoma, an oligodendroglioma, an ependymoma, or a combination thereof.   The method of claim 2, wherein the brain tumor is a polymorphic glioblastoma.   The method of claim 2, wherein the brain tumor is a metastatic brain tumor.   5. The method according to any one of claims 1-4, further comprising administering an additional therapeutic agent.   6. The method of claim 5, wherein the additional therapeutic agent is temozolomide.   7. The method of any one of claims 1-6, further comprising subjecting the subject to radiation therapy.   The method according to claim 1, wherein the brain tumor is characterized by the expression of a mutant form of KRAS.   A method for inhibiting the growth of brain tumor cells, comprising contacting the brain tumor cells with plinabulin.   10. The method of claim 9, wherein the contacting comprises administering an effective amount of plinabulin to a subject having the brain tumor cells.   A method for inducing apoptosis of brain tumor cells, comprising contacting the brain tumor cells with plinabulin.   12. The method of claim 11, wherein the contacting comprises administering an effective amount of plinabulin to a subject having the brain tumor cells.   A method for inhibiting progression of brain tumor, comprising administering an effective amount of plinabulin to a subject in need thereof.